Thursday, April 18, 2013
Applications Thursday: PHERAstar FS enhances cell based fluorescence assays
BMG LABTECH application note 206 describes the performance of the PHERAstar FS in detection of intracellular calcium and dopamine receptor activation. This is performed using two Invitrogen assays: The Fluo-4 Direct Calcium Assay and the Tango D1-bla U2OS GPCR Cellular Assay; which is based on arrestin recruitment to the dopamine receptor. The performance in each assay highlights the qualities of the PHERAstar FS that make it such a fantastic HTS microplate reader.
Both assays use the bottom reading capabilities of the PHERAstar FS which is highly desirable for performance of cell based assays such as these. Furthermore the calcium assay exploits the injection at point of measurement capability of the PHERAstar FS which allows for precise measurement of fast kinetic assay such as this.
Finally, the GPCR assay employs the dual emission detection of the PHERAstar FS to assess whether FRET is occurring for substrate labeled with coumarin and fluorescein, which is indicative of inactivity of the receptor. Upon receptor activation beta-lactamase is expressed which cleaves the substrate leading to a decrease in FRET. Using the dual emission detection capabilities you can monitor the FRET emission at 530 nm and emission at 460 nm indicative of loss of FRET simultaneously.
For more information on this application and the many other applications that BMG LABTECH microplate readers can perform please visit the Applications Center on the BMG website.